FACE™ Maker

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The FACE™ Method

In FACE, cells are cultured in 96-well plates and stimulated to induce the pathway of interest. Following stimulation, the cells are fixed rapidly, which preserves activation-specific protein modifications. Each well is then incubated with a primary antibody specific for the activated protein of interest. Subsequent incubation with secondary HRP-conjugated antibody and developing solution provides a colorimetric or chemiluminescent readout that is quantitative and reproducible (Figure 1). The number of cells in each well can be normalized easily with the provided Crystal Violet solution. FACE Kits also contain primary antibody specific for the native inactive protein, so you can monitor both native and activated protein levels in the same experiment. FACE eliminates cellular extractions, radioactive kinase assays, time-consuming Westerns and inefficient epitope interactions that occur on membranes. FACE is a highly sensitive high-throughput assay designed for detecting activated proteins within mammalian cells.

Figure 1: Flow chart of the FACE process.

Choose your own antibodies with FACE Maker

FACE Maker Kits differ from other FACE Kits in that they do not include any antibodies. Instead, they provide all other components for running a FACE assay, enabling you to study any phospho-protein you choose. So, you must supply a phospho-specific antibody that detects only the phosphorylated form of your protein of interest. You may also run a parallel experiment with an antibody that recognizes the total protein of interest regardless of its modification state to compare the effects of treatment conditions versus a basal state. You will also need to supply species-appropriate HRP-conjugated secondary antibodies for simple detection.